应用PCR和FISH方法诊断X-连锁隐性鱼鳞病3例  

Three patients with X-linked recessive ichthyosis to be diagnosed through PCR and FISH techniques

作　　者：张学奇[1] 刘晶晶[1] 林孝华[1] 邵笑红[1] 蔡剑峰[1] 徐云升[1] 李智铭[1] 

机构地区：[1]温州医学院附属第一医院皮肤科,温州325000

出　　处：《安徽医科大学学报》2016年第12期1859-1862,共4页Acta Universitatis Medicinalis Anhui

基　　金：温州科学技术局课题(编号:Y20140576);国家自然科学基金(编号:81272987)

摘　　要：应用多聚酶链反应(PCR)和荧光原位杂交(FISH)方法对3例X-连锁隐性鱼鳞病(XLRI)患者进行分子诊断。对2个X-连锁隐性鱼鳞病家系3例患者及家族成员采集外周血标本后,先采用PCR扩增STS基因的两末端序列及侧翼的微卫星标记;再应用c DNA STS基因特异性探针对间期白细胞核进行FISH检测。选取X染色体上自DXS1139至DXF22S1区域内7个微卫星标记进行PCR扩增以确定DNA缺失大小。PCR检测STS基因两端序列结果没有扩增产物。FISH方法也证实了患者STS基因DNA序列存在缺失。应用多态性微卫星序列进行PCR扩增,结果表明患者存在X染色体上STS基因及其侧翼序列,自DXS1139至DXF22S1区域内,DNA缺失达1.97 Mb。PCR和FISH方法证实XLRI患者STS基因及其两侧翼序列DNA缺失,具有重要的遗传咨询意义。To diagnose patients of X-linked recessive ichthyosis（ XLRI） involving a complete deletion of the steroid sulfatase（ STS） gene detected with PCR and FISH. Cutaneous examination was performed and peripheral blood samples were collected from three patients affected XLRI. PCR amplification of both ends of the STS gene was first performed,and then FISH in metaphase leukocytes was performed with the c DNA STS probe. Finally,amplification of the regional markers from DXS1139 to DXF22S1 on the X chromosome was performed through PCR. PCR amplification both the ends of STS gene using genomic DNA failed to produce any amplicons. However,each exon was successfully amplified using genomic DNA and primers from control samples. Subsequently,FISH analyses showed no red signal for the STS gene and only one green signal for the X-chromosome centromere in three patients.Analyses of the deletion of STS gene and its flanking sequences in one Chinese pedigree with XLRI using polymerase chain reaction and fluorescence in situ hybridization techniques were performed,which has great significance for the genetic counseling.

关 键 词：X连锁隐性鱼鳞病 STS基因 STS基因缺失 PCR FISH 

分 类 号：R758.25[医药卫生—皮肤病学与性病学] Q33[医药卫生—临床医学]